The ITGB8 Knockout HT29 Polyclonal Cells are a CRISPR/Cas9-mediated gene-disrupted polyclonal cell population with functional disruption of ITGB8, encoding integrin subunit beta-8. This knockout model provides a physiologically relevant platform for loss-of-function studies, maintaining population heterogeneity while eliminating target gene expression. The polyclonal format avoids clonal artifacts and offers a robust tool for investigating ITGB8-dependent mechanisms in a human colorectal adenocarcinoma background.
The HT29 cell line is a well-characterized human colorectal adenocarcinoma model with epithelial morphology, widely used in cancer biology and drug screening. Derived from a primary colon adenocarcinoma, HT29 cells retain intestinal epithelial features and harbor mutations in APC, TP53, and KRAS, making them valuable for studying colorectal cancer progression, EMT, and tumor microenvironment interactions.
ITGB8 pairs with ITGAV to form the ??v??8 integrin, which binds the RGD motif of the LAP of TGF-?? and promotes activation of latent TGF-??. Active TGF-?? engages TGFBR1/TGFBR2, phosphorylating SMAD2 and SMAD3, which complex with SMAD4 to regulate transcription of EMT genes like SNAI1 and SNAI2. Non-canonical pathways involve FAK, SRC, AKT, and MAPK. Upstream regulators include ECM proteins (fibronectin, vitronectin), ITGAV, TGF-??, and mechanical cues. Interacting factors such as thrombospondin-1 further modulate this pathway, positioning ITGB8 as a key mediator linking ECM cues to TGF-??-driven cell adhesion, migration, and invasion.
In HT29 cells, ITGB8-mediated TGF-?? activation promotes EMT, tumor invasion, and metastasis. Disrupting ITGB8 allows dissection of ??v??8 integrin??s contribution to TGF-?? signaling independently of other integrins. The knockout population permits assessment of how loss of ITGB8 affects SMAD phosphorylation, mesenchymal marker expression, and invasive behavior while retaining tumor cell heterogeneity, thus aiding mechanistic studies of colorectal cancer progression and therapeutic targeting of the ??v??8/TGF-?? axis.
Research applications encompass TGF-?? signaling studies, EMT and metastasis research, drug target validation, and fibrosis models. Representative assays include Western blotting for ITGB8 and phospho-SMAD2/3, RT-qPCR for SNAI1/2, Transwell migration/invasion, flow cytometry for integrin expression, TGF-?? luciferase reporter assays, and immunofluorescence for integrin localization. The polyclonal knockout cells are suitable for high-throughput screening and mechanistic studies. For technical details, please contact Ascent Research.
